基因重组鲨鱼软骨血管生成抑制因子的发酵条件优化

The optimization of the recombinant Shark Cartilage Angiogenesis Inhibiting Factor's fermentation conditions

采用三角摇瓶来优化基因工程菌表达重组鲨鱼软骨血管生成抑制因子(SCAIF)的最佳发酵条件,并以此为基础在Biotop CF-10L自动控制发酵罐进行发酵培养.基因工程菌BL21(DE3)/pET3c(SCAIF)诱导表达SCAIF蛋白的最佳摇瓶培养条件是:接种量为2%、装液量为75 mL/500 mL、加入IPTG的诱导时间为培养2 h后、IPTG浓度为0.25 mmol/L、诱导时间为2 h.BL21(DE3)/pET3c(SCAIF)菌种在Biotop CF-10L自动控制发酵罐中的条件为:以10%的接种量接种到10 L发酵培养基中,设定pH7.0,温度37℃,培养4 h后,加入终浓度为0.5 mmol/L的IPTG,诱导2 h后终止发酵.发酵罐中获得的菌体量为10.2 g/L,蛋白表达率为25%左右.

To determine the optimal condition for the expression of recombinant Shark Cartilage Angiogenesis Inhibiting Factor （ SCAIF）, IPTG concentration, induction time, harvest period, Inoculum concentration, liquid volume were optimized in this study in flask. It was found that higher expression of rSCAIF could be achieved under the following conditions： 2% inoculum was added to 75 mL/500 mL LB medium. After 2 h of culture, 0. 25 mmol/L IPTG was added to induce rSCAIF expression. The recombinant BL21 （ DE3 ）/SCAIF was harvested after 2 h of induction. Large scale gene expression was performed in Biotop CF-10L fermenter. Started with 10% inoculation, the cell was cultured in pH7.0 fermentation medium for 4hours, followed by induction with 0. 5 mmol/L IPTG for 3 hours. The final biomass was 10. 2 g/L, and the expressed rSCAIF was about 25% of total cellular protein.